2015-2018年辽宁省脊灰实验室细胞敏感性监测与评价
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摘要
目的评价辽宁省脊灰实验室2015-2018年所用细胞系的质量控制情况,为急性驰缓性麻痹(acute flaccid paralysis)病例监测提供可靠的质量保证。方法以辽宁省脊灰实验室2013年制备的脊灰病毒Ⅰ、Ⅱ、Ⅲ型质量控制(Laboratory quality control standard,LQC)株为标准毒株,转脊灰病毒受体的小鼠肺细胞(L20B细胞)和横纹肌肉瘤细胞细胞(RD细胞)由中国疾病预防控制中心脊灰实验室提供。支原体的检测用PCR方法,细胞敏感性检测用标准毒株滴定法。结果辽宁省脊灰实验室2015-2018年使用的L20B和RD细胞系支原体检测均为阴性,两种细胞系的病毒滴度与LQC参考值比较波动幅度均在±0.5 log CCID50/0.1 ml以内。结论 2015-2018年辽宁省脊灰实验室使用的细胞系状态稳定,无支原体污染,对脊灰病毒敏感性良好,为脊灰实验室网络运转和辽宁省AFP病例监测提供了准确、可靠的病毒学依据。
Objectives To evaluate the quality control of cell lines used in Liaoning Polio laboratory from 2015 to 2018 and provide reliable quality assurance for acute flaccid paralysis(AFP)monitoring. Methods The LQC of poliovirus I,II and III was made in 2013 in Liaoning Province,and the L20 B and RD cell lines were provided by the National Polio Laboratory. Mycoplasma was detected by PCR and cell sensitivity was tested with standard strain titration. Results The L20 B and RD cell lines used in Liaoning Polio Laboratory from September 2015 to September 2018 were negative for mycoplasman; the viral titers of the two cell lines fluctuated within ±0.5 logCCID50/0.1 ml compared with the value of LQC. Conclusion The cell lines used in Liaoning Polio Laboratory from 2015 to 2018 were stable. Cell lines were free of mycoplasma and sensitive to poliovirus. It provides an accurate and reliable virological basis for the network operation of polio laboratory and AFP case monitoring in Liaoning Province.
Objectives To evaluate the quality control of cell lines used in Liaoning Polio laboratory from 2015 to 2018 and provide reliable quality assurance for acute flaccid paralysis(AFP)monitoring. Methods The LQC of poliovirus I,II and III was made in 2013 in Liaoning Province,and the L20 B and RD cell lines were provided by the National Polio Laboratory. Mycoplasma was detected by PCR and cell sensitivity was tested with standard strain titration. Results The L20 B and RD cell lines used in Liaoning Polio Laboratory from September 2015 to September 2018 were negative for mycoplasman; the viral titers of the two cell lines fluctuated within ±0.5 logCCID50/0.1 ml compared with the value of LQC. Conclusion The cell lines used in Liaoning Polio Laboratory from 2015 to 2018 were stable. Cell lines were free of mycoplasma and sensitive to poliovirus. It provides an accurate and reliable virological basis for the network operation of polio laboratory and AFP case monitoring in Liaoning Province.
引文
[1]王东艳,张勇,安洪秋,等.中国脊髓灰质炎实验室网络2007~2015年细胞敏感性监测与评价[J].中国疫苗和免疫,2016,2(22):138-142.
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[5]祝双利,李晓嫘,朱晖,等.中国2015年脊髓灰质炎实验室网络的运转与评价[J].中国疫苗和免疫,2017,23(2):127-133.
[6]陈玫,赵娜,张俊棉,等.2006-2011年河北省脊髓灰质炎实验室细胞敏感性监测结果分析[J].预防医学情报杂志,2013,29(7):569-571.
[7]苟东东.浅谈细胞规模化培养过程中引起污染的类型及防治策略探讨[J].科技与创新,2018(12):39-41.
[8]武昱孜,张旭,华利忠,等.支原体对细胞培养污染的研究概况[J].动物医学进展,2013,34(9):112-117.
[9]WHO. Outbreak of poliomyelitis, Republic of the Congo, September 2010-February 2011[J]. Weekly Epidemiological Record,2011.
[10]Wang HB,Zhu SL,Zheng JS,et al. Sero-Survey of polio antibodies during wild poliovirus outbreak in Souther Xinjiang Uygur Autonomous region, China[J]. PLOS ONE,2014,9(7):1-9.
[11]段亚彤,袁伟,任丽萍,等.辽宁省2006-2011年健康人群脊灰抗体水平分析[J].中国公共卫生,2013,29(10):1545-1546.
[12]王东艳,祝双利,张勇,等.中国脊髓灰质炎实验室网络开展细胞系支原体检测及现状分析[J].中国疫苗和免疫,2017,23(2):8-12.
[13]夏晴.细胞库支原体检测体系以及支原体去除方法的建立[D].上海:上海交通大学,2014.
[2]WHO. Polio laboratory manual[M]. 4thedition. Geneva:WHO,2004:73-80.
[3]黄海军,高其双,彭霞,等.细胞支原体巢式PCR检测方法的建立及应用[J].湖北农业科学,2014,53(3):690-693.
[4]唐海淑,崔惠,翟啸虎,等.新疆维吾尔自治区脊髓灰质炎实验室2014-2017年细胞系质量控制评价[J].中国疫苗和免疫,2018,24(2):57-60.
[5]祝双利,李晓嫘,朱晖,等.中国2015年脊髓灰质炎实验室网络的运转与评价[J].中国疫苗和免疫,2017,23(2):127-133.
[6]陈玫,赵娜,张俊棉,等.2006-2011年河北省脊髓灰质炎实验室细胞敏感性监测结果分析[J].预防医学情报杂志,2013,29(7):569-571.
[7]苟东东.浅谈细胞规模化培养过程中引起污染的类型及防治策略探讨[J].科技与创新,2018(12):39-41.
[8]武昱孜,张旭,华利忠,等.支原体对细胞培养污染的研究概况[J].动物医学进展,2013,34(9):112-117.
[9]WHO. Outbreak of poliomyelitis, Republic of the Congo, September 2010-February 2011[J]. Weekly Epidemiological Record,2011.
[10]Wang HB,Zhu SL,Zheng JS,et al. Sero-Survey of polio antibodies during wild poliovirus outbreak in Souther Xinjiang Uygur Autonomous region, China[J]. PLOS ONE,2014,9(7):1-9.
[11]段亚彤,袁伟,任丽萍,等.辽宁省2006-2011年健康人群脊灰抗体水平分析[J].中国公共卫生,2013,29(10):1545-1546.
[12]王东艳,祝双利,张勇,等.中国脊髓灰质炎实验室网络开展细胞系支原体检测及现状分析[J].中国疫苗和免疫,2017,23(2):8-12.
[13]夏晴.细胞库支原体检测体系以及支原体去除方法的建立[D].上海:上海交通大学,2014.